Dear Phillip,
I'm quite conscious on the SOLiD mate pair library, the steps and the structure of the mate (p1-insert-mpl-mpr-insert-p2) .. it's about one year I'm trying to improve the robustness of the method. Belong to my experience the protocol of the mp isn't satisfactory and mainly replicable.
SO, I'm trying to find other way to overcame some steps that in my opinion are limiting. The circularization is one of these points. The problem to circularize DNA is that the IA (in the 5500 xl is formed by mpr and mpl) contains part of the ligation site for the annealing in sequencing. Therefore, the importance of knowing the IA sequence ( and it is irreplaceable).
I was interested in that post, because you (or someone else) wrote the IA sequence with a string of NNNNN. I thought that my sanger sequence on the IA was with a double signal (2 picks) because of the NNNNN string.
I'm not sure if it's more clear my question on the Mpl Mpr sequence!
thanks for your answers
chiara
I'm quite conscious on the SOLiD mate pair library, the steps and the structure of the mate (p1-insert-mpl-mpr-insert-p2) .. it's about one year I'm trying to improve the robustness of the method. Belong to my experience the protocol of the mp isn't satisfactory and mainly replicable.
SO, I'm trying to find other way to overcame some steps that in my opinion are limiting. The circularization is one of these points. The problem to circularize DNA is that the IA (in the 5500 xl is formed by mpr and mpl) contains part of the ligation site for the annealing in sequencing. Therefore, the importance of knowing the IA sequence ( and it is irreplaceable).
I was interested in that post, because you (or someone else) wrote the IA sequence with a string of NNNNN. I thought that my sanger sequence on the IA was with a double signal (2 picks) because of the NNNNN string.
I'm not sure if it's more clear my question on the Mpl Mpr sequence!
thanks for your answers
chiara
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